Supplemental Information. Spatial Auxin Signaling. Controls Leaf Flattening in Arabidopsis
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1 Current Biology, Volume 27 Supplemental Information Spatial Auxin Signaling Controls Leaf Flattening in Arabidopsis Chunmei Guan, Binbin Wu, Ting Yu, Qingqing Wang, Naden T. Krogan, Xigang Liu, and Yuling Jiao
2 Figure S1. Different markers expressed in young leaf primordia. Related to Figure 1. (A) P2 and P3 leaf primordia of Arabidopsis. S, stipule. Scale bar, 50 m. (B) Expression patterns of ARF6, ARF7, ARF8 and ARF19 in the leaf primordia. Transverse sections through parf6::n3gfp, pnph4::n3gfp, parf8::n3gfp, and parf19::n3gfp SAM and leaf primordia region. GFP signals are shown in green and PI staining in red. Scale bars, 20 m. (C) Expression patterns of MP, DR5, and DII in the leaf primordia. Longitudinal and transverse sections through pmp::mp-gfp, pdr5::gfper, and p35s::dii-venus SAM and leaf primordia region. GFP and venus signals are shown in green. Scale bars, 20 m. (D) Optical section analysis of young leaf primordia expressing pdr5v2::ntdtomato and pdr5::n3gfp. DR5, green. DR5v2, magenta. Purple dotted lines indicate transverse sections. Scale bars, 20 m.
3 Figure S2. Dof5.8 expression level and seedling phenotypes of pmp::mp -EAR-GR transgenic plants. Related to Figure 2. (A) Dof5.8 expression under Dex treatment in pmp::mp -EAR-GR transgenic plants. Data are presented as mean ± SD for more than three independent experiments. *P < (B) Phenotypes of pmp::mp -EAR-GR transgenic plants under 10 m Dex treatment at different developmental stages. Scale bars, 1 mm and 10 mm for the upper and lower panels, respectively.
4 Figure S3. Auxin induces WOX1 expression and the overexpression of MP promotes WOX1 adaxial expression. Related to Figure 3. (A) SlWOX1 expression 24 h after IAA treatment in tomato P 1 leaf primordia. Data are presented as mean ± SD for more than three independent experiments. *P < (B) Serial sections of pmp::mp leaf primordia in ISH analysis of WOX1 expression pattern. P 4 in the right section is comparable to P 4 of Col-0 in (A) along the proximodistal axis. Ladders colored in yellow indicate cells with WOX1 transcripts. Scale bars, 20 m.
5 Figure S4. Seedling phenotype of pwox1::wox1 and pwox1 ::WOX1 transgenic plants in wox1-2 prs double mutant background. Related to Figure 4. Scale bars, 1 mm for 9 d and 12 d, and 10 mm for 30 d.
6 Figure S5. WOX1 and PRS expression pattern in arf3-1 arf4-2 and arf2-6 arf3-1 arf4-2 mutants and the seedling phenotypes of the triple mutants. Related to Figure 5. (A) Serial sections of arf3-1 arf4-2 leaf primordia in ISH analysis of WOX1 expression pattern. M, meristem. Scale bars, 20 m. (B) Seedling phenotypes of arf3-1 arf4-2 double and arf2-6 arf3-1 arf4-2 triple mutants. White arrows indicate typical rosette leaves with different phenotypes in double and triple mutants respectively. Scale bars, 1 mm for 9 d, and 10 mm for others. (C) A trumpet-like leaf of arf2-6 arf3-1 arf4-2 triple mutant. Scale bar, 1 mm.
7 Figure S6. Phenotypes of p35s::amir-arf transgenic plants. Related to Figure 5. (A) Vegetative phenotypes of p35s::amir-arf plants. Scale bar, 10 mm. (B) Leaf abaxial side phenotypes of wild-type and p35s::amir-arf rosette leaves. Scale bar, 1 mm. (C) RT-qPCR analysis of ARF2, ARF3, and ARF4 expression levels in p35s::amir-arf rosette leaves.
8 Table S1. Primers used for RT-PCR, ChIP-PCR and Y1H. Related to STAR Methods. Primer Primer sequence (5-3 ) WOX1-RT-F WOX1-RT-R PRS-RT-F PRS-RT-R ARF2-RT-F ARF2-RT-R ARF3-RT-F ARF3-RT-R ARF4-RT-F ARF4-RT-R MP-RT-F MP-RT-R ACTIN2F ACTIN2R WOX1-1-ChIP-F WOX1-1-ChIP-R WOX1-2-ChIP-F WOX1-2-ChIP-R WOX1-3-ChIP-F WOX1-3-ChIP-R WOX1-4-ChIP-F WOX1-4-ChIP-R PRS-1-ChIP-F PRS-1-ChIP-R PRS-2-ChIP-F PRS-2-ChIP-R PRS-3-ChIP-F PRS-3-ChIP-R PRS-4-ChIP-F PRS-4-ChIP-R WOX1-1-Y1H-F WOX1-1-Y1H-R WOX1-2-Y1H-F WOX1-2-Y1H-R WOX1-3-Y1H-F WOX1-3-Y1H-R WOX1-4-Y1H-F WOX1-4-Y1H-R PRS-1-Y1H-F PRS-1-Y1H-R CTGGATATGTTCGGTCGGATG CTCCACCCGTATATTCGCTG TGTCCTTTGATTGCTGCTCTC TCTTCAGCTCCACTTTTGGTGCAG GCGAGTTCGGAGGTTTCAATGAAA TCTGTAAAGAGCAGCCTCAGGGTCC CGCCTACTCAATAACCGATCATC ACGGCCCACACCAAATGTT CGCTTAAATCATTCCCGCAAT ACTTGTTGGCTTGGTAAGCAAAG GTTGAAAGACCAGTCAGGTAC ATGTCTCTTTGGTTGCCCTC GAGAGGTTACATGTTCACCACAAC GTGAACGATTCCTGGACCTGCCTC CTGAGCTCGACAGAAAAGGGGGATTTAA GACTCGAGCATTCCTCCCATCTCTCCCC GCTGATCACTGCATTTATTG GTCTCGAGCCCAACCAAATATATGTCAC CCCATAAGCCAAAATAGCCA GGAGAAGGGAGAGAGACAGCG CTGAGCTCCGAGAACGCCAGAAACGACG CACTCGAGCTCTGGTTGCGTGTCGCATC GATTATTTCTAGAAGAGGAC AGAATCTCCAGAAACTGAGC CAGACAATTCTATGCCTGAT CTTTACATGGACAGACAAGC GTTAATGAGTACTGGCGTTT GGAGAGAAAGAGAAAAGGAAC CTCTTGCGTCCCTTTCCAAT AGGACTCATTCTCCGTTCAGAC CTGAGCTCGACAGAAAAGGGGGATTTAA GACTCGAGCATTCCTCCCATCTCTCCCC GAGAGCTCTCACGTCGTTATAAGCTCCT GTCTCGAGCCCAACCAAATATATGTCAC GAGAGCTCGGTATCTCTTTCTCTCTCTC CACTCGAGGGCTTTGTGGTTTTGGAACC CTGAGCTCCGAGAACGCCAGAAACGACG CACTCGAGCTCTGGTTGCGTGTCGCATC CGGAGCTCGATTATTTCTAGAAGAGGAC CGGTCGACAGAATCTCCAGAAACTGAGC
9 PRS-2-Y1H-F PRS-2-Y1H-R PRS-3-Y1H-F PRS-3-Y1H-R PRS-4-Y1H-F PRS-4-Y1H-R ARF4-Y1H-F ARF4-Y1H-R GCGAGCTCCAGACAATTCTATGCCTGAT GCGTCGACCTTTACATGGACAGACAAGC GCGAGCTCGTTAATGAGTACTGGCGTTT GCGTCGACGGAGAGAAAGAGAAAAGGAAC GCGAGCTCCTCTTGCGTCCCTTTCCAAT GCGTCGACAGGACTCATTCTCCGTTCAGAC AAAGCGGCCGCCATGGAATTTGACTTGAATAC TTGGCGCGCCAACCCTAGTGATTGTAGGAG
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